Extended Data Fig. 5: Long duration activator binding correlates with long duration SAGA binding.
From: Single-molecule analysis of transcription activation: dynamics of SAGA coactivator recruitment
a. Dual color rastergrams of Gal4-SNAPDY649-VP16 and Spt7-SNAPDY549 binding events on template with five Gal4 binding sites. Activator binding records from 100 randomly chosen non-chromatinized (upper panels) and chromatinized (lower panel) template locations were binary-coded (red color indicates colocalization with DNA) and sorted by time to first activator binding, from bottom to top. Next, corresponding binary-coded SAGA binding records (green) from the same template were overlaid. Simultaneous binding of SAGA and activator was colored as blue. Increasing activator concentrations (3, 10, and 30 nM Gal4-SNAPDY649-VP16) are shown from left to right. b. Gal4-SNAPDY649-VP16 (10 nM) binding to non-chromatinized DNA templates containing a single Gal4 binding site (1xG4bs), in the absence of NTPs. Left panel shows cumulative time-to-initial-binding distribution of Gal4-SNAPDY649-VP16 on the template (red) or off-target (gray) locations. Dashed lines are curve fits (parameters in Supplementary Table 5). Right panel displays dual color rastergram of Gal4-SNAPDY649-VP16 activator (red) and Spt7-SNAPDY549 (green) binding records from 100 randomly chosen templates. Intervals when both are bound are colored in blue. c. Long duration activator occupancy can occur on DNA with a single Gal4 binding site. Survival plots of Gal4-SNAPDY649-VP16 occupancy intervals at locations of single Gal4 UAS containing template (red) or at off-target locations (gray). Bi-exponential fit parameters and numbers of events are listed in Supplementary Table 6.
